HOW DID OUR ANTIBODY PERFORM?

ANTIBODY CUSTOMER REVIEW: Phospho-ErbB-3 (Y1197) Polyclonal Antibody (STJ90642 )

What was used?Primary antibody: STJ90642 Phospho-ErbB-3 (Y1197)

Provider: St John's Laboratory

Dilution ratio: 1:1000

Application: Western Blot

Materials for validation: MCF7 cell line

Click for product data sheet PDF

What Was The Protocol?Treatment of Materials When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors

(Sigma) and equal volume of 2X loading buffer were added for 10 minutes at 95˚C.

Gel Electrophoresis 10% Polyacrylamide gel with a constant voltage of 160V for 60 minutes.

Transfer 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100V for 75 minutes.

Blocking 1X TBST with 5% BSA for 1 hour.

Primary Antibody Probing Primary antibody was added to the solution of 1X TBST with 5% BSA, and left at 4˚C overnight.

Membrane Wash Membrane was washed with 1X TBST 3 times.

Secondary Antibody Probing Secondary antibody was added to the solution of 1X TBST with 5% BSA with a 1:2500 dilution ratio, for 1 hour.

Membrane Wash Membrane was washed with 1X TBST 3 times.

Visualization ECL visualization for 3 min (the visualization time was determined by actual result. It was ensured that the visualization time was identical for the same antigen).

What was the protocol?Gel electrophoresis information: 10% polyacrylamide gel, constant voltage of 160V for 60 minutes.

Transfer information: 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100V for 75 minutes.

No. Antigen Loading amount

Primary antibody

Primary antibody

dilution ratio

Secondaryantibody

dilution ratioTarget band KD Visualization

time

1 MCF7 15µg

STJ96177Phospho-ErbB-3

(Y1197) antibody

1:1000 1:2500 ~170KD 40S

What were the results?

What did the customer think?

Antibody Specificity:

Antibody RPPA Rating:

Testing results were provided independently by R. Bernardo at the Royal College of Surgeons in Ireland. They stated “In MCF7 cell line, there were target bands detected by Phospho-ErbB-3 (Y1197) STJ90642 Antibody.”

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